Tn5 Transposase production

Tn5 transposase is the key enzyme in forming transposomes for random transposon insertions. It is sold by Epicentre at ridiculously high price. Here, we make it from a plasmid provided by Prof. Wolfgang Hillen PMID 16820464. The protocol also builds on the NEB IMPACT-CN protein purification kit.

Materials

pWH1891 plasmid (kind gift of Prof. Wolfgang Hillen)

BL21(DE3)pLysS cells

TEGX buffer

10 mM Tris-HCl pH 7.5

700 mM NaCl

1 mM EDTA

10% glycerol

0.1% Triton X-100

Storage buffer (Epicentre)

50% glycerol

50 mM Tris-HCl pH 7.5

100 mM NaCl

0.1 mM EDTA -- our version has 5 mM EDTA, since we never want in-vitro action